应用QF-PCR检测先心病患儿22q11.2微缺失方法的建立

Research of establishing a way of detecting the 22q11.2 microdeletion with QF-PCR in patients with congenital heart disease

目的:探讨运用荧光定量PCR方法检测先心病患儿22号染色体q11.2微缺失的准确度及灵敏度,在此基础上,建立一种准确度高、特异性强、快速、高通量、费用低且适合在国内开展的诊断22q11.2微缺失的方法。方法:对84例CHD患儿进行外周血染色体检查,以染色体核型正常的先心病患儿TBX1基因附近SNP位点为靶位点采用QF-PCR方法检测22q11.2的微缺失,将其结果与FISH结果进行比较,同时选取30例正常儿童作为对照。结果:84例CHD患儿中有2例染色体异常,其余82例患儿中,有5例QF-PCR方法显示存在22q11.2微缺失,与FISH检测结果完全一致,而正常对照组均未见异常。结论:QF-PCR方法检测先心病患儿22q11.2微缺失准确度高、特异性强,与传统的FISH方法比较,具有快速、简便、高通量的特点,可以作为22号染色体微缺失的诊断依据,为明确CHD患儿病因提供可靠的实验室依据。

Objective： To determine whether quantitative fluorescence polymerase chain reaction （ QF - PCR） could be used to de- tect the microdeletion of 22ql 1.2 in patients with congenital heart disease and to establish a rapid, inexpensive, sensitive, specific and high throughout method for screening 22ql 1. 2DS. Methods： The traditional peripheral venous chromosomal karyotypes of 84 CHD children were examined. The SNP site which located in upstream of TBX1 gene on 22 chromosome as a target site was chosen. QF - PCR and FISH Were used to screen 22ql 1.2 mierodeletion in normal karyotypes children respectively. At the same time, 30 healthy children were chosen as controls. Results ： Among the 84 test samples, 2 were abnormal karyotypes. In the other 82 test samples with normal karyotypes, 5 were de- lermined to the microdeletion of 22ql 1.2 using QF - PCR. And the result was similar with the ones with FISH. There was no microdeletion in control samples using QF - PCR and FISH. Conclusion ： QF - PCR is a sensitive and specific way to detect the microdeletion of 22ql 1.2 in patients with congenital heart disease. It also has the advantages of rapid, inexpensive and high throughout compared With the conventional gold standard method - FISH. It can be used as a new screening way for 22ql 1.2DS, and providing reliable laboratory evidence for identifica- tion of CHD